1. A comparative study of the bispecific monoclonal antibody, blinatumomab expression in CHO cells and E. coli.
- Author
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Naddafi F, Shirazi FH, Talebkhan Y, Tabarzad M, Barkhordari F, Aliabadi Farahani Z, Bayat E, Moazzami R, Mahboudi F, and Davami F
- Subjects
- Animals, CHO Cells, Cricetulus, Antibodies, Bispecific biosynthesis, Antibodies, Bispecific chemistry, Antibodies, Bispecific isolation & purification, Escherichia coli genetics, Escherichia coli metabolism, Gene Expression
- Abstract
The "bispecifics" market improved over the past decade due to the development of many technological platforms including bispecific T cell engagers (BiTEs). The approval of blinatumomab, the most advanced bispecific T-cell engager (BiTE) in clinical trials, can be a significant milestone in the development of bispecific antibodies. Both Chinese hamster ovary (CHO) cells and E. coli strain are considered as the most widely used hosts for the large-scale production of therapeutic monoclonal antibodies. Since both of the economic and qualitative aspects of protein production are important in industry, selection of a suitable protein expression system is very critical. The BsAb gene was cloned into the expression vectors FC550A-1, pcDNA3.1 (+), and PET22b and 6 × His-tagged BsAb then purified on a Ni-NTA chromatography column. Both SDS-PAGE and Western blotting analysis of the purified protein demonstrated that blinatumomab was successfully expressed as a 55 kDa in both expression systems. The antigen-binding properties of blinatumomab were compared in the mammalian system versus Escherichia coli. The results showed that the purified antibody from a mammalian expression system has better binding activity than the one from E. coli host.
- Published
- 2018
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